1.CD40-ligand down-regulates inhibitor of differentiation 3 expression in WEHI-231 B lymphoma cells
Journal of Medical Postgraduates 2004;0(02):-
Objective: To study mouse Id3(mId3) protein expression in WEHI-231 B lymphoma cells after CD40 ligand(CD40L) treatment. Methods: The mId3 cDNA was amplified from total RNA of WEHI-231 cells by RT-PCR and introduced into the p3FLAG-CMV-10 vector.The 3FLAG-Id3 fusion gene was amplified from p3FLAG-Id3-CMV-10 vector and subcloned into retroviral vector pMX-CITE/IRES-EGFP.Recombinant retrovirus was produced by transient transfection of the Phoenix-ecotropic packaging cell line with pMX-3FLAG-Id3-EGFP vector using FuGene-6 transfection reagent.(WEHI)-231 cells were transduced with recombinant retrovirus.EGFP expression was monitored by flow cytometry.Two cell lines stably expressing 3FLAG-Id3 fusion protein were generated. Results: The proportion of EGFP-positive cells in the population transduced with pMX-3FLAG-EGFP remained constant up to 2 days after transduction,whereas the proportion in the population of pMX-3FLAG-Id3-EGFP-transduced cells decreased with time.After coculturing with NIH3T3 cells or medium alone,the proportion of EGFP-positive cells in pMX-3FLAG-Id3-EGFP-transduced WEHI-231 cells decreased with time,whereas after coculturing with CD40L-NIH3T3 cells,the proportion of EGFP-positive cells in the transduced cells remained constant up to 2 days,suggesting that CD40L could rescue WEHI-231 from Id3 overexpressioninduced growth arrest.After coculturing with CD40L/NIH3T3 or NIH3T3 cells,3FLAG-Id3 fusion protein stably expressing WEHI-231 cells were harvested,lysed,and immunoblotted by using anti-FLAG antibody.The results showed that treatment via CD40 ligation induced significant downregulation of Id3 expression in pMX3FLAG-Id3-EGFP tansduced WEHI 231 cells. Conclusion:CD40 ligation pvevents the Id3 induced cell growth avvest by the dousn-regulation of Id3 expression.
2.Cell morphology observation of breast cancer on fine needle aspiration biopsy
Chunlin FU ; Xianghong LING ; Junchuan LI ; Yu ZHANG
Journal of International Oncology 2011;38(4):310-315
Objective To summarize the cytological features of breast carcinoma by fine needle aspiration and differential diagonsis of breast hyperplasia and breast flbroadenoma. Methods Cytological features about 175 cases of breast cancer were analyzed,contrasted with 76 cases of breast hyperplasia and 93 eases of fibroadenoma. And 20 cases of low-diagnosis breast cancer were analyzed, compared with 13 cases of excessive diagnosis. Results The followings were found:disorderly arranged cells in 169 cases of breast cancer, about 96.9% in 175 cases,loose distance between the nucleus in 125 cases of all cases (about 71.4%). The percentage of small groups or scattered cells, medium size cells, round nucleus and irregular border nucleus was 34.3% (60/175) ,81.1% (142/175), 88.6% (155/175) and 28.6% (50/175) respectively. The nucleus/cytoplasm ratio was increased in 105 cases of all (about 60%). The nucleus/cytoplasm ratio was obviously reduced in 15 cases of all (about 8.6%). Big nucleolis were observed in 49 cases of all (about 28%). There was significant difference between nucleus size in 136 cases of all (about 77.7%). There was not myoepithelial cell in 168 cases of all (about 96%). The percentage of visible necrosis, scsttered round nucleus and integrity of cytoplasmic was 13.7% (24/175) ,89.7% (157/175), 66.9% (117/175) respectively. The cases of all above features were significantly more than those of breast hyperplasia and breast fibroadenoma (P <0.05). The percentage of large number of cells was 52.6% in 175 breast cancer cases,higher than 5.3% in 76 breast hyperplasia cases. There was a significant difference(P <0.05) between two groups. In 20 cases of low-diagnosis breast cancer the percentage of round nucleus with integrity of cytoplasmic, special arrangement cells was 70% (14 cases) ,35% (7 cases) ,higher than the cases in 13 excessive diagnosis cases. There was a significant difference(P < 0.05) between two groups. Conclusion The cell morphological characteristics of breast caner by fine needle aspiration are large number of cells, loose distance between the nucleus, disorderly arranged cells, small guoups or scattered cells, integrity of cytoplasmic, increased nuclear, round nuclear, irregular border nucleus,increased or decreased nucleus/cytoplasm ratio, big nucleoli, significant difference in nuclear size, no myoepithelial cell, visible necrosis. Most of breast cancer can be distinguished from breast hyperplasia and breast fibroadenoma by above characteristics. All those cytological characteristics are helpful to identify difficult case.
3.Expression and significance ofβ-catenin in gastric adenocarcinoma and the metastasis of lymphnode
Tianxing DU ; Jiyuan YANG ; Long LI ; Huichuan ZHAO ; Tingxuan CHEN ; Junchuan LI
Journal of International Oncology 2014;(9):696-699
Objective To explore the expressions ofβ-catenin in gastric adenocarcinoma primary tumors and metastatic lymph nodes,and to determine if it is associated with infiltration degree and whether it can provide the basis for gastric cancer metastasis.Methods The expressions ofβ-catenin were detected in 156 patients with gastric adenocarcinoma specimens,40 cases with the corresponding lymph nodes and 12 cases of normal gastric tissues by the method of immunohistochemistry.The expressions ofβ-catenin in gastric adenocarcinoma and corre-sponding lymph node metastases were analysed.The relationship between the expression of β-catenin and the clinical pathological features of gastric adenocarcinoma was ascertained,and difference between them was observed.Results The positive expression rate ofβ-catenin in normal gastric tissue membrane was 100.0%(12/12).The expression rate ofβ-catenin was 29.5%(46/156)in adenocarcinoma cell membrane,and it was 10.0%(4/40)in metastatic lymph nodes cell membrane.The positive expression ofβ-catenin was not associated with patient′s age(χ2 =2.160,P=0.142),gender (χ2 =1.229,P=0.268),but it was associated with tumor infiltration degree (χ2 =4.032,P=0.045 ),degree of tumor differentiation (χ2 =6.093,P=0.048),tumor stage (χ2 =4.591,P=0.032),and metastasis lymph nodes (χ2 =4.485,P=0.034).The incidence of abnor-mal or loss expression ofβ-catenin in gastric adenocarcinoma metastatic lymph nodes was higer than that in gastric adenocarcinoma (χ2 =6.362,P=0.012).Conclusion The expression of β-catenin will be a great help to judge the biological behaviors such as the malignant degree of tumor and the capacity of tumor invasion and metas-tasis.
4.Expression of Oct4 and Wnt2 in human glioma tissues and its clinical significance
Guangrui ZHAO ; Mingcan WU ; Shijie CHEN ; Junchuan LI ; Tingxuan CHEN ; Yuan YAO
Chinese Journal of Cancer Biotherapy 2009;16(6):629-632
Objective:To investigate the expression of Oct4 and Wnt2 in human glioma tissues and its relationship with the clinicopathological features of glioma. Methods: Fifty-six paraffin blocks were obtained from glioma patients receiving surgery. The diagnosis of these patients were confirmed by pathology in our hospital from 2006-2009. Immunohistochemi-cal staining was used to examine Oct4 and Wnt2 expression in the brain tissues of 10 patients with acute brain injury and 56 glioma tissues (including 15 recurrent cases). Results: The normal brain tissues were negative of Oct4, with only one case showing weak Wnt2 expression. Thirty-four of the 56 glioma tissues showed positive expression of Oct4 (60.7%), and 40 showed positive expression of Wnt2 (71.4%). Positive expression rates of Oct4 and Wnt2 in low-grade and high-grade glioma tissues were 46.2 %, 73.3% and 57.7 %, 83.3%, respectively (P < 0.05). Oct4 positive rates in the recrudescence and newly diagnosed glioma tissues were 86.7% and 51.2%, respectively (P < 0.05). Oct4 expression in the glioma tissues was positively correlated with that of Wnt2 (r = 0.537, P < 0.01). Conclusion: Expression of Oct4 and Wnt2 is associated with the malignant degrees of glioma, and Oct4 expression is related to the recurrence of glioma. Oct4 might participate in the development and progression of brain glioma through Wnt signaling pathway.
5.Relationship between TGF-betal and BRCA2 expression and clinical related factors in breast cancer
Junchuan LI ; Hongyu ZHU ; Tingxuan CHEN ; Guihua DAI ; Lanying ZOU ; Huichuan ZHAO
Journal of Chinese Physician 2009;11(3):344-346
Objective To explore the relationship between TGF-beta1,BRCA2,HER2,ER,PR and clinical factors in breast cancer.Methods The expression of TGF-beta1,BRCA2,HEB2,ER,PB in 67 cases breast carcinoma were detected by immunohistochemistry staining SP method.The correlation of the results with other parameters which included age,pathohistological grade,status of auxiliary lymph nodes were analyzed by mono-factor unconditional logistic regression analysis.Results The expression of TGF-beta1 was correlated with BRCA2.and the expression of BRCA2 was correlated with TGF-beta1 and c-erbB-2 in breast cancer.Condusion Overexpression of BRCA2 was related with TGF-beta1 and HER2 in breast carcinoma.It was useful in breast carcinoma prognosis by detecting these three factors.